Yeast gain-of-function mutations reveal structure-function relationships conserved among different subfamilies of transient receptor potential channels

Transient receptor potential (TRP) channels found in animals, protists, and fungi are primary chemo-, thermo-, or mechanosensors. Current research emphasizes the characteristics of individual channels in each animal TRP subfamily but not the mechanisms common across subfamilies. A forward genetic screen of the TrpY1, the yeast TRP channel, recovered gain-of-function (GOF) mutations with phenotype in vivo and in vitro. Single-channel patch-clamp analyses of these GOF-mutant channels show prominent aberrations in open probability and channel kinetics. These mutations revealed functionally important aromatic amino acid residues in four locations: at the intracellular end of the fifth transmembrane helix (TM5), at both ends of TM6, and at the immediate extension of TM6. These aromatics have counterparts in most TRP subfamilies. The one in TM5 (F380L) aligns precisely with an exceptional Drosophila mutant allele (F550I) that causes constitutive activity in the canonical TRP channel, resulting in rapid and severe retinal degeneration beyond mere loss of phototaxis. Thus, this phenylalanine maintains the balance of various functional states (conformations) of a channel for insect phototransduction as well as one for fungal mechanotransduction. This residue is among a small cluster of phenylalanines found in all known subfamilies of TRP channels. This unique case illustrates that GOF mutations can reveal structure-function principles that can be generalized across different TRP subfamilies. It appears that the conserved aromatics in the four locations have conserved functions in most TRP channels. The possible mechanistic roles of these aromatics and the further use of yeast genetics to dissect TRP channels are discussed.     

Correction of proton resonance frequency shift temperature maps for magnetic field disturbances using fat signal

PURPOSE: To improve the immunity of the proton resonance frequency shift (PRFS) method of MRI temperature mapping against magnetic field disturbances. Since PRFS is a phase-sensitive method, it misinterprets magnetic field disturbances as artifact temperature changes. If not corrected, the resulting temperature artifacts can completely obscure the true temperature estimation, especially if the temperature elevations are small. MATERIALS AND METHODS: Since the fat protons experience the same magnetic field disturbances as the water protons, but no temperature-related frequency shift, the fat signal has been used for correcting PRFS temperature maps for the disturbances. A simple correction method is proposed that has either better compensation capability than the phase correction methods previously reported or higher spatial and temporal resolution than the spectroscopic correction methods previously reported. The evaluated method is based on the utilization of several gradient and spin echoes acquired within one repetition interval with water- and fat-selective scans. RESULTS: In a series of phantom experiments, the improved method is shown to enable the reconstruction of accurate temperature maps in spite of interscan motion, suboptimal fat-water separation, and a wide range of magnetic field disturbances. CONCLUSION: Our approach can be used for the guidance of thermal therapies involving tissues containing fat or surrounded by fat.     

The contribution to the epidemiology of gastrointestinal nematodes of sheep with special focus on the survival of infective larvae in winter conditions

Tracer tests conducted over a 3-year period were aimed at measuring the level and species nematode composition of survival on pastures with a special focus on winter months. The survival of infective larvae in chilly conditions is not significantly affected by Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus and Chabertia ovina. On the contrary, the number of Teladorsagia circumcincta and Nematodirus filicollis significantly increased in milder winter conditions. The results confirmed an epidemiological strategy of overwintering in the arrested stage for Teladorsagia circumcincta and Nematodirus filicollis; the epidemiological strategy of genus Trichostrongylus used both strategies—in particular the tolerance of free-living stages to cold conditions. Part of the population overwintered in the arrested stage as well.     

The association between alcohol use, alcohol use disorders and tuberculosis (TB). A systematic review

Background  

In 2004, tuberculosis (TB) was responsible for 2.5% of global mortality (among men 3.1%; among women 1.8%) and 2.2% of global burden of disease (men 2.7%; women 1.7%). The present work portrays accumulated evidence on the association between alcohol consumption and TB with the aim to clarify the nature of the relationship.     

Platelets Strongly Induce Hepatocyte Proliferation with IGF-1 and HGF In Vitro

BACKGROUND: It is well known that platelets have a thrombotic effect. However, platelets play an important role not only in hemostasis but also in wound healing and tissue regeneration. Platelets have been reported to accumulate in the liver and promote liver regeneration after an extended hepatectomy, but the mechanism is unclear. The present study was designed to clarify the mechanism by which platelets have a direct proliferative effect on hepatocytes in vitro. MATERIALS AND METHODS: Hepatocytes obtained from male BALB/c mice by collagenase digestion and immortalized hepatocytes (TLR2) were used. To elucidate the mechanism of the proliferative effect of platelets, DNA synthesis of hepatocytes was measured under various conditions and the related cellular signals were analyzed. Chromatographic analysis was also performed to clarify which elements of platelets have mitogenic activity. RESULTS: DNA synthesis significantly increased in the hepatocytes cultured with platelets (P < 0.001). However, when the platelets and hepatocytes were separated, the platelets did not have a proliferative effect. Whole disrupted platelets, the supernatant fraction, and fresh isolated platelets had a similar proliferative effect, while the membrane fraction did not. After the addition of platelets, both Akt and extracellular signal-regulated kinases ERK1/2 were activated, but extracellular signal-regulated kinase STAT3 was not activated. Some mitogenic fractions were obtained from the platelet extracts by gel exclusion chromatography; the fractions were rich in hepatocyte growth factor and IGF-1. CONCLUSIONS: Direct contact between platelets and hepatocytes was necessary for the proliferative effect. The direct contact initiated signal transduction involved in growth factor activation. Hepatocyte growth factor, vascular endothelial growth factor, and insulin-like growth factor-1, rather than platelet-derived growth factor, mainly contributed to hepatocyte proliferation.     

Bedroom air quality and vacuuming frequency are associated with repeat child asthma hospital admissions

Objective: Indoor environment factors have been associated with risk of asthma exacerbations in children but little is known about their role on asthma hospital readmissions. As children in Western societies continually spend more time indoors, understanding the influence of these factors on asthma exacerbation is important. We examined the role of indoor environmental and lifestyle characteristics on child asthma readmissions. Methods: A hospital-based case–control study recruited 22 children readmitted for asthma and 22 controls not readmitted for asthma. Logistic regression models were used to examine the association between aeroallergens and fungi in the bedroom and indoor lifestyle characteristics factors for asthma readmissions. To determine the best possible set of predictors among a large set of risk factors, we used random forests (RF) techniques. Results: Higher levels of airborne Cladosporium and yeast in the child’s bedroom increased risk of readmission (OR?=?1.68, 95% CI 1.04–2.72 and OR?=?1.52, 95% CI 0.99–2.34, respectively). Carpeted floors in the bedroom and synthetic doonas were also associated with increase in asthma readmissions (OR?=?4.07, 95% CI 1.03–16.06 and OR?=?14.6, 95% CI 1.26–169.4, respectively). In the home, frequent vacuuming using bagged cleaners increased risk of asthma readmission OR?=?15.7 (95% CI 2.82–87.2). Conclusions: Factors in the child’s bedroom play an important role in increasing the risk of asthma hospital readmissions. These findings have major clinical implications as the identified potential risk factors may be modifiable. Further epidemiological studies with larger samples are necessary to evaluate these associations further.     

Group testing in mediation analysis

We consider the scenario where there is an exposure, multiple biologically defined sets of biomarkers, and an outcome. We propose a new two-step procedure that tests if any of the sets of biomarkers mediate the exposure/outcome relationship, while maintaining a prespecified familywise error rate. The first step of the proposed procedure is a screening step that removes all groups that are unlikely to be strongly associated with both the exposure and the outcome. The second step adapts recent advances in postselection inference to test if there are true mediators in each of the remaining candidate sets. We use simulation to show that this simple two-step procedure has higher statistical power to detect true mediating sets when compared with existing procedures. We then use our two-step procedure to identify a set of Lysine-related metabolites that potentially mediate the known relationship between increased body mass index and the increased risk of estrogen-receptor positive breast cancer in postmenopausal women.